21029747
not annotated - annotated - LINNAEUS only
A fluorescence resonance energy transfer-based fluorometer assay for screening anti-coxsackievirus B3 compounds.
In view of the need to develop a simple and rapid method to screen for antiviral therapeutic agents, a fluorescence resonance energy transfer (FRET)-based reporter system consisting of engineered mammalian cells expressing a cyan fluorescent protein-yellow fluorescent protein (CFP-YFP) pair linked by a short peptide containing the cleavage site of viral protease 2A (2A(pro)) was developed. By detecting the 2A(pro) produced early during the virus infection cycle, the CFP-YFP pair effectively identifies infectious coxsackievirus B3 (CVB3), a picornavirus that causes viral myocarditis in humans. The reporter system was used to screen a library of 2000 drugs and natural products for potential antiviral compounds. The reporter cells were treated with the test compounds, challenged with CVB3, and then examined using a fluorometer at 24h post-infection. Sixty-four compounds, mostly therapeutic drugs, antimicrobial compounds and compounds with unknown functions, caused at least 50% inhibition of 2A(pro) activity. Three known antiviral compounds, cosmosiin, ribavirin and baicalein, were also identified in the screening. The developed method is an effective strategy for rapid screening, and identifies compounds that inhibit CVB3 2A(pro). This method should be a valuable aid in the antiviral drug discovery effort.
Ann file
T1 Out-of-scope 84 101 coxsackievirus B3
N1 Reference T1 Taxonomy:138949
T2 Species 633 650 coxsackievirus B3
N2 Reference T2 Taxonomy:138949
T3 Species 652 656 CVB3
N3 Reference T3 Taxonomy:138949
T4 Species 707 713 humans
N4 Reference T4 Taxonomy:9606
T5 Species 907 911 CVB3
N5 Reference T5 Taxonomy:138949
T6 Species 1349 1353 CVB3
N6 Reference T6 Taxonomy:138949